Phytocannabinoids in the treatment of cancer

ABSTRACT

This invention relates to the use of phytocannabinoids, either in an isolated form or in the form of a botanical drug substance (BDS) in the treatment of cancer. Preferably the cancer to be treated is cancer of the prostate, cancer of the breast or cancer of the colon.

RELATED APPLICATIONS

This application is a national stage filing under 35 U.S.C. §371 ofinternational application PCT/GB2011/050487, filed Mar. 11, 2011, whichwas published under PCT Article 21(2) in English.

This invention relates to the use of phytocannabinoids, either in anisolated form or in the form of a botanical drug substance (BDS), as aprophylactic or in the treatment of cancer. Typically the cancer to betreated is a cancer of the: prostate, breast, skin, glioma, colon, lungor a bone or lymph metastasis. The phytocannabinoids may be used incombination with other cancer treatments.

BACKGROUND

Cancer is a class of diseases which occurs because cells becomeimmortalised; they fail to heed customary signals to turn off growthwhich is a normal function of remodelling in the body that requirescells to die on cue. Apoptosis, or programmed cell death, can becomedefective and when this happens malignant transformation can take place.The immortalised cells grow beyond their normal limits and invadeadjacent tissues. The malignant cells may also metastasise and spread toother locations in the body via the bloodstream or lymphatic system.Cancer cells often form a mass known as a tumour.

There are about 200 different types of cancer; the cancers can start inany type of body tissue although many cancers will metastasise intoother body tissues. There are many different causes of cancer and theseinclude; carcinogens, age, genetic mutations, immune system problems,diet, weight, lifestyle, environmental factors such as pollutants, someviruses for example the human papilloma virus (HPV) is implicated incervical cancer and some bacterial infections are also known to causecancers.

There are many different treatment options for cancer and the treatmentsought is often determined by the type and stage of the cancer.Treatment options include; chemotherapeutic drug treatment, hormonaldrug treatment, radiotherapy, surgery, complementary therapies andcombinations thereof.

Prostate cancer is the most common type of cancer in men and accountsfor 24% of all UK male cancers. In 2006 there were over 35,000 new casesof prostate cancer diagnosed in the UK alone.

The prostate is a gland in the male reproductive system and symptoms ofcancer in the prostate can include pain, difficulty urinating, problemswith sexual intercourse and erectile dysfunction. Prostate cancer maymetastasise to the bones and or lymph nodes. Treatment options forprostate cancer include surgery, radiation therapy, chemotherapy andhormone treatment.

Hormone treatment usually involves treatment with an anti-androgen suchas cyproterone acetate, flutamide or bicalutamide, either alone or incombination with a chemotherapeutic agent. These treatments work to stopthe production of testosterone (androgen) which can slow down tumourgrowth or even shrink the tumour. While the prostate cancer cells areresponding to anti-androgens, they are referred to as‘hormone-sensitive’ prostate cancer. Unfortunately, after a few years oftreatment with anti-androgens the prostate cancer stops responding tohormone treatment and is termed ‘hormone-insensitive’ prostate cancer.At this stage the cancer growth cannot be controlled by the hormonetreatment.

In order to test the effectiveness of different compounds in thetreatment of either hormone-sensitive or hormone-insensitive prostatecancer two different cell lines can be used. The cell line LNCaP arehormone-sensitive prostate cancer cells which were derived from asupraclavicular lymph node metastasis in a 50 year old male in 1977. Thecell line DU-145 are hormone-insensitive prostate cancer cells whichwere derived from a brain metastasis.

It is known that expression levels of both cannabinoid receptors, CB1and CB2, were significantly higher in CA-human papillomavirus-10(virally transformed cells derived from adenocarcinoma of human prostatetissue), and other human prostate cells LNCaP, DU-145, PC3, and CWR22RN1than in human prostate epithelial and PZ-HPV-7 (virally transformedcells derived from normal human prostate tissue) cells (Sarfaraz, 2005).

Additionally it is known that WIN-55,212-2 (mixed CB1/CB2 agonist)treatment with hormone sensitive LNCaP cells resulted in a dose—(1-10Mmol/L) and time-dependent (24-48 hours) inhibition of cell growth.Blocking of CB1 and CB2 receptors by their antagonists SR141716 (CB1)and SR144528 (CB2) significantly prevented this effect.

These results suggested that WIN-55,212-2 or other cannabinoid receptoragonists could be developed as novel therapeutic agents for thetreatment of prostate cancer.

Cannabis has been ascribed to be both a carcinogen and anti-canceragent. In particular smoking cannabis is known to be carcinogenic as thecannabis smoke contains at least 50 different known carcinogeniccompounds, many of which are the same substances found in smokedtobacco. One of these carcinogens, benzopyrene is known to cause canceras it alters a gene called p53, which is a tumour suppressor gene.Cannabis contains the substance tetrahydrocannabinol (THC) which hasbeen shown to cause benzopyrene to promote the p53 gene to change.

Researchers however have discovered that some cannabinoids, includingTHC and cannabidiol (CBD) are able to promote the re-emergence ofapoptosis so that some tumours will heed the signals, stop dividing, anddie. The process of apoptosis is judged by observation of severalphenomena including: reduced cellular volume, condensation of nuclearchromatin, changes in distribution of phospholipids in plasma membranephospholipids, and cleavage of chromatin into DNA fragments called DNAladders.

Another method by which tumours grow is by ensuring that they arenourished: they send out signals to promote angiogenesis, the growth ofnew blood vessels. Cannabinoids may turn off these signals as well.

Cannabinoids have been shown to have an anti-proliferative effect ondifferent cancer cell lines. The cannabinoids THC, THCA, CBD, CBDA, CBGand CBC and the cannabinoid BDS THC and CBD were tested on eightdifferent cell lines including DU-145 (hormone-sensitive prostatecancer), MDA-MB-231 (breast cancer), CaCo-2 (colorectal cancer) and C6(glioma cells). The data for each cannabinoid in each different type ofcancer varied but generally the best data were observed with CBD or CBDBDS. The IC50 values for all the cannabinoids on the DU-145 were quitehigh inferring that none of the cannabinoids tested were particularlyeffective in the inhibition of hormone-insensitive prostate cancer(Ligresti, 2006).

Several transient receptor potential (TRP) channels have been implicatedin the survival, growth and spread of prostate and other cancers. TRPM8is expressed in sensory neurons, where it responds to cold and tocooling agents, notably menthol, but it is also abundantly expressed inthe prostate. In particular TRPM8 is over-expressed in hormone-sensitiveprostate cancer cells, but expression of TRPM8 is almost completelyablated once the cancer becomes hormone-insensitive and in patientsreceiving anti-androgen therapy. Expression of TRPM8 is stimulated byandrogens in hormone-sensitive prostate cancer cell lines (LNCaP). Thereis evidence that expression of TRPM8 is required for survival ofprostate cancer cells.

The mechanism of such an action of TRPM8 is likely to relate to itsability to modulate intracellular calcium, and possibly even thedistribution of calcium within the cell. The latter point may beimportant because of the localisation of TRPM8 in the prostate cancercell. While found on the cell membrane, it is also found on theendoplasmic reticulum; thus any potential therapeutic agent whichtargets the TRPM8 receptor must be able to gain good access to theintracellular space.

The endogenous cannabinoid anandamide has been shown to antagonise TRPM8(De Petrocellis, 2007). The authors also showed that stimulation of CB1receptors transiently antagonised TRPM8 receptors expressed on the samecells.

The application WO 2008/129258 describes the use ofcannabinoid-containing plant extracts in the prevention or treatment ofdiseases or conditions that are alleviated by blockade of one or moretypes of TRP channel. Different binding potentials of thecannabinoid-containing plant extracts at the TRPA1 and TRPM8 channelsare described. The diseases and conditions to be prevented or treatedinclude: neuropathic pain, inflammation, vasoconstriction or cancer.

The TRPM8 receptor has also been found in breast, colon and skincancers.

It has been shown that CBD is able to able to down-regulate theexpression of the DNA binding protein inhibitor, Id-1 in human breastcancer cells (McAllister, 2007). The CBD concentrations effective atinhibiting Id-1 expression correlated with those used to inhibit theproliferative and invasive phenotype of breast cancer cells. CBD wasable to inhibit Id-1 expression at the mRNA and protein level in aconcentration-dependent fashion.

CBD has also been shown to inhibit human cancer cell proliferation andinvasion through differential modulation of the ERK and ROS pathways,and that sustained activation of the ERK pathway leads todown-regulation of Id-1 expression. It was also demonstrated that CBDup-regulates the pro-differentiation agent, Id-2. Using a mouse 4T1 cellline and a model of metastatic breast cancer, CBD significantly reducedmetastatic spread. As such CBD may represent a promising treatment ofbreast cancer in patients with secondary tumours (McAllister, 2009).

Recent evidence indicates that CBD is a GPR55 antagonist; this raisesthe possibility that this receptor may underlie the effects of CBD onbreast and other tumour cells. GPR55 couples to G12/13 and thedownstream activation of the RhoA, rac1 and cdc42 small GTPases; thispathway is crucial in cytoskeletal reorganisation and cell migration.Increased G12/13 expression has been found in early stage human breastcancer cells taken by biopsy and inhibition of G13 decreases the levelof breast cancer cell metastasis in vivo (Kelly et al, 2007).

The anti-proliferative effects of CBD have also been evaluated on U87and U373 human glioma cell lines, (Massi, 2004). The anti-proliferativeeffect of CBD was correlated to induction of apoptosis, as determined bycytofluorimetric analysis and single-strand DNA staining, which was notreverted by cannabinoid antagonists. In addition CBD, administered s.c.to nude mice at the dose of 0.5 mg/mouse, significantly inhibited thegrowth of subcutaneously implanted U87 human glioma cells. It wasconcluded that CBD was able to produce a significant anti-tumouractivity both in vitro and in vivo, thus suggesting a possibleapplication of CBD as a chemotherapeutic agent.

The application WO/2006/037981 describes the use of the cannabinoid CBDto prevent tumour cells migrating or metastisising from an area ofuncontrolled growth to an area away from the original tumour site. CBDcaused a concentration-dependent inhibition of the migration of U87glioma cells, quantified in a Boyden chamber. Since these cells expressboth cannabinoid CB1 and CB2 receptors in the membrane, the group alsoevaluated their engagement in the anti-migratory effect of CBD.

Cannabinoids have been shown to play a fundamental role in the controlof cell survival/cell death. It has been reported that cannabinoids mayinduce proliferation, growth arrest, or apoptosis in a number of cells,including neurons, lymphocytes, and various transformed neural andnon-neural cells, and that cannabinoids induce apoptosis of glioma cellsin culture and regression of malignant gliomas in vivo (Guzman, 2001).

A pilot clinical study of THC in patients with recurrent glioblastomamultiforme has been conducted. This pilot phase I trial consisted ofnine patients with recurrent glioblastoma multiforme who wereadministered THC intra-tumourally. The patients had previously failedstandard therapy (surgery and radiotherapy) and had clear evidence oftumour progression. The primary end point of the study was to determinethe safety of intracranial THC administration. They also evaluated THCaction on the length of survival and various tumour-cell parameters.Median survival of the cohort from the beginning of cannabinoidadministration was 24 weeks (95% confidence interval: 15-33).

The application WO 2008/144475 describes treating cell proliferationdisorders including cancer with cannabidiol derivatives either alone orin combination with THC or a derivative thereof.

The application WO 03/063847 describes the use of CBDA or CBDVA as anactive pharmaceutical substance. The focus of the application provides atreatment for nausea, vomiting emesis and motion sickness.

The application WO 2009/147439 describes the use of a combination ofcannabinoids, particularly tetrahydrocannabinol (THC) and cannabidiol(CBD), in the manufacture of a medicament for use in the treatment ofcancer. In particular the cancer to be treated is a brain tumour, moreparticularly a glioma; more particularly still a glioblastoma multiforme(GBM).

The application WO 2009/147438 describes the use of one or morecannabinoids, particularly THC and/or CBD in combination with anon-cannabinoid chemotherapeutic agent in the manufacture of amedicament for use in the treatment of cancer. In particular the cancerto be treated is a brain tumour, more particularly a glioma, moreparticularly still a glioblastoma multiforme (GBM). The non-cannabinoidchemotherapeutic agent may be a selective estrogen receptor modulator oran alkylating agent.

The literature and corresponding patent applications demonstrate thegeneral usefulness of cannabinoids in the area of cancer research andtreatment.

It is an object of the present invention to find improved and/oralternative cancer therapies. To this end a platform of datarepresenting the use of isolated phytocannabinoids and phytocannabinoidbotanical drug substances (BDS) in different aspects of the treatment ofcancer is provided and the results extrapolated to identify groups ofphytocannabinoids, whether isolated or in the form of a BDS, whichappear more promising than others in specific treatments.

DEFINITIONS AND ABBREVIATIONS

Definitions of some of the terms used to describe the invention aredetailed below:

The phytocannabinoids described in the present application are listedbelow along with their standard abbreviations.

CBC Cannabichromene

CBCV Cannabichromenic acid

CBD Cannabidiol

CBDA Cannabidiolic acid

CBDV Cannabidivarin

CBG Cannabigerol

CBGV Cannabigerol propyl variant

CBL Cannabicyclol

CBN Cannabinol

CBNV Cannabinol propyl variant

CBO Cannabitriol

THC Tetrahydrocannabinol

THCA Tetrahydrocannabinolic acid

THCV Tetrahydrocannabivarin

THCVA Tetrahydrocannabivarinic acid

The table above is not exhaustive and merely details the cannabinoidswhich are identified in the present application for reference. So farover 60 different cannabinoids have been identified and thesecannabinoids can be split into different groups as follows:Phytocannabinoids; Endocannabinoids and Synthetic cannabinoids.

“Phytocannabinoids” are cannabinoids that originate from nature and canbe found in the cannabis plant. The phytocannabinoids can be isolatedcannabinoids or present as a botanical drug substance.

An “isolated cannabinoid” is defined as a phytocannabinoid that has beenextracted from the cannabis plant and purified to such an extent thatall the additional components such as secondary and minor cannabinoidsand the non-cannabinoid fraction have been removed.

A “botanical drug substance” or “BDS” is defined in the Guidance forIndustry Botanical Drug Products Draft Guidance, August 2000, USDepartment of Health and Human Services, Food and Drug AdministrationCentre for Drug Evaluation and Research as: “A drug derived from one ormore plants, algae, or microscopic fungi. It is prepared from botanicalraw materials by one or more of the following processes: pulverisation,decoction, expression, aqueous extraction, ethanolic extraction or othersimilar processes.” A botanical drug substance does not include a highlypurified or chemically modified substance derived from natural sources.Thus, in the case of cannabis, BDS derived from cannabis plants do notinclude highly purified Pharmacopoeial grade cannabinoids.

“Endocannabinoids” are the cannabinoids that are produced endogenouslyby human or animal bodies. Up or down regulation of the endocannabinoidsystem may be useful in the treatment of some diseases or conditions.

“Synthetic cannabinoids” are compounds that have a cannabinoid-likestructure yet are manufactured using chemical means. Depending on themethod of manufacture the synthetic cannabinoid may comprise a racemicmixture of cannabinoids, in contrast to an isolated cannabinoid whichwill be a single enantiomer.

Phytocannabinoids can be found as either the neutral (decarboxylatedform) or the carboxylic acid form depending on the method used toextract the cannabinoids. For example it is known that heating thecarboxylic acid form will cause most of the carboxylic acid form todecarboxylate into the neutral form.

Phytocannabinoids can also occur as either the pentyl (5 carbon atoms)or propyl (3 carbon atoms) variant. Initially it was thought that thepropyl and pentyl variants would have similar properties, however recentresearch has found that this may not be true. For example thephytocannabinoid THC is known to be a CB1 receptor agonist whereas thepropyl variant THCV has been discovered to be a CB1 receptor antagonistmeaning that it has almost opposite effects.

In the present invention a BDS is considered to have two components: thephytocannabinoid-containing component and the non-phytocannabinoidcontaining component. Preferably the phytocannabinoid-containingcomponent is the larger component comprising greater than 50% (w/w) ofthe total BDS and the non-phytocannabinoid containing component is thesmaller component comprising less than 50% (w/w) of the total BDS.

The amount of phytocannabinoid-containing component in the BDS may begreater than 55%, through 60%, 65%, 70%, 75%, 80% to 85% or more of thetotal extract. The actual amount is likely to depend on the startingmaterial used and the method of extraction used.

The “principle phytocannabinoid” in a BDS is the phytocannabinoid thatis present in an amount that is higher than that of the otherphytocannabinoids. Preferably the principle phytocannabinoid is presentin an amount greater than 40% (w/w) of the total extract. Morepreferably the principle phytocannabinoid is present in an amountgreater than 50% (w/w) of the total extract. More preferably still theprinciple phytocannabinoid is present in an amount greater than 60%(w/w) of the total extract.

The amount of the principle phytocannabinoid in the BDS is preferablygreater than 75% of the phytocannabinoid-containing fraction, morepreferably still greater than 85% of the phytocannabinoid-containingfraction, and more preferably still greater than 95% of thephytocannabinoid-containing fraction.

In some cases, such as where the principle cannabinoid is either CBDV orTHCVA the amount of the principle phytocannabinoid in the BDS is lower.Here the amount of phytocannabinoid is preferably greater than 55% ofthe phytocannabinoid-containing fraction.

The “secondary phytocannabinoid/s” in a BDS is the phytocannabinoid/sthat is/are present in significant proportions. Preferably the secondaryphytocannabinoid is present in an amount greater than 5% (w/w) of thetotal extract, more preferably greater than 10% (w/w) of the totalextract, more preferably still greater than 15% (w/w) of the totalextract. Some BDS's will have two or more secondary phytocannabinoidsthat are present in significant amounts. However not all BDS's will havea secondary phytocannabinoid. For example CBG BDS does not have asecondary phytocannabinoid in its extract.

The “minor phytocannabinoid/s” in a BDS can be described as theremainder of all the phytocannabinoid components once the principle andsecondary phytocannabinoids are accounted for. Preferably the minorphytocannabinoids are present in total in an amount of less than 10%(w/w) of the total extract, more preferably still less than 5% (w/w) ofthe total extract, and most preferably the minor phytocannabinoid ispresent in an amount less than 2% (w/w) of the total extract.

Typically the non-phytocannabinoid containing component of the BDScomprises terpenes, sterols, triglycerides, alkanes, squalenes,tocopherols and carotenoids.

These compounds may play an important role in the pharmacology of theBDS either alone or in combination with the phytocannabinoid.

The “terpene fraction” may be of significance and can be broken down bythe type of terpene: monoterpene or sesquiterpene. These terpenecomponents can be further defined in a similar manner to thecannabinoids.

The amount of non-phytocannabinoid containing component in the BDS maybe less than 45%, through 40%, 35%, 30%, 25%, 20% to 15% or less of thetotal extract. The actual amount is likely to depend on the startingmaterial used and the method of extraction used.

The “principle monoterpene/s” in a BDS is the monoterpene that ispresent in an amount that is higher than that of the other monoterpenes.Preferably the principle monoterpene/s is present in an amount greaterthan 20% (w/w) of the total terpene content. More preferably theprinciple monoterpene is present in an amount greater than 30% (w/w) ofthe total terpene content, more preferably still greater than 40% (w/w)of the total terpene content, and more preferably still greater than 50%(w/w) of the total terpene content. The principle monoterpene ispreferably a myrcene or pinene. In some cases there may be two principlemonoterpenes. Where this is the case the principle monoterpenes arepreferably a pinene and/or a myrcene.

The “principle sesquiterpene” in a BDS is the sesquiterpene that ispresent in an amount that is higher than all the other terpenes.Preferably the principle sesquiterpene is present in an amount greaterthan 20% (w/w) of the total terpene content, more preferably still tgreater than 30% (w/w) of the total terpene content. The principlesesquiterpene is preferably a caryophyllene and/or a humulene.

The sesquiterpene components may have a “secondary sesquiterpene”. Thesecondary monoterpene is preferably a pinene, which is preferablypresent at an amount greater than 5% (w/w) of the total terpene content,more preferably the secondary terpene is present at an amount greaterthan 10% (w/w) of the total terpene content.

The secondary sesquiterpene is preferably a humulene which is preferablypresent at an amount greater than 5% (w/w) of the total terpene content,more preferably the secondary terpene is present at an amount greaterthan 10% (w/w) of the total terpene content.

Alternatively botanical extracts may be prepared by introducing isolatedphytocannabinoids into a non-cannabinoid plant fraction as can beobtained from a zero cannabinoid plant or a CBG-free BDS.

BRIEF SUMMARY OF THE DISCLOSURE

In accordance with a first aspect of the present invention there isprovided a cannabis plant extract comprising a phytocannabinoidcontaining component and a non-phytocannabinoid containing component,for use in medicine, wherein the phytocannabinoid containing componentcomprises at least 50% (w/w) of the cannabis plant extract and thenon-phytocannabinoid containing component comprises a monoterpenefraction and a sesquiterpene fraction, in which a principle monoterpenesub-fraction is selected from myrcenes or pinenes and a principlesesquiterpene sub-fraction is selected from caryophyllenes or humulenes.

In accordance with a second aspect of the present invention there isprovided the use of a cannabis plant extract comprising aphytocannabinoid containing component and a non-phytocannabinoidcontaining component, for use in the manufacture of a medicament for usein medicine, wherein the phytocannabinoid containing component comprisesat least 50% (w/w) of the cannabis plant extract and thenon-phytocannabinoid containing component comprises a monoterpenefraction and a sesquiterpene fraction, in which a principle monoterpenesub-fraction is selected from myrcenes or pinenes and a principlesesquiterpene sub-fraction is selected from caryophyllenes or humulenes.

In accordance with a third aspect of the present invention there isprovided a method of treating a patient comprising administering atherapeutically effective amount of a cannabis plant extract comprisinga phytocannabinoid containing component and a non-phytocannabinoidcontaining component, wherein the phytocannabinoid containing componentcomprises at least 50% (w/w) of the cannabis plant extract and thenon-phytocannabinoid containing component comprises a monoterpenefraction and a sesquiterpene fraction, in which a principle monoterpenesub-fraction is selected from myrcenes or pinenes and a principlesesquiterpene sub-fraction is selected from caryophyllenes or humulenesto the patient.

Preferably principle monoterpene sub-fraction comprises myrcenes and thesecondary monoterpene sub-fraction comprises pinenes. In anotherembodiment the principle monoterpene sub-fraction are both myrcenes andpinenes.

Preferably the principle sesquiterpene sub-fraction comprisescaryophyllenes and secondary sesquiterpene sub-fraction compriseshumulenes.

Preferably the principle phytocannabinoid is selected from the groupconsisting of: THCV, CBDV, CBGV, THCVA, THCA, CBDA, CBG, THC, CBD andCBC.

Preferably the non-phytocannabinoid containing component furthercomprises one or more compounds from the group consisting of:diterpenes; triterpenes; sterols; triglycerides; alkanes; squalenes;tocopherols; and carotenoids.

In one embodiment the cannabis plant extract comprises the principlephytocannabinoid CBG and the phytocannabinoid containing componentcomprises 61-75% (w/w) of the cannabis plant extract. Preferably theextract further comprises greater than 88% (w/w) CBG of the totalphytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid THC and the phytocannabinoid containingcomponent comprises 77-94% (w/w) of the cannabis plant extract.Preferably the extract further comprises 78-95% (w/w) THC of the totalphytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid CBD and the phytocannabinoid containingcomponent comprises 76-96% (w/w) of the cannabis plant extract.Preferably the extract further comprises 72-88% (w/w) CBD of the totalphytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid CBC and the phytocannabinoid containingcomponent comprises 49-60% (w/w) of the cannabis plant extract.Preferably the extract further comprises 71-87% (w/w) CBC of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoids CBD and CBL. More preferably still theCBD comprises 6.5-8% (w/w) of the total phytocannabinoid fraction andthe CBL comprises 5.8-7.1 (w/w) of the total phytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid THCV and the phytocannabinoid containingcomponent comprises 74-90% (w/w) of the cannabis plant extract.Preferably the extract further comprises 71-87% (w/w) THCV of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoid THC. More preferably still the THCcomprises 14.8-18% (w/w) of the total phytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid CBDV and the phytocannabinoid containingcomponent comprises 64-78% (w/w) of the cannabis plant extract.Preferably the extract further comprises 52-64% (w/w) CBDV of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoids CBD and CBCV. More preferably still theCBD comprises 22.4-27.4% (w/w) of the total phytocannabinoid fractionand the CBCV comprises 5.5-6.7 (w/w) of the total phytocannabinoidfraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid CBGV and the phytocannabinoid containingcomponent comprises 54-66% (w/w) of the cannabis plant extract.Preferably the extract further comprises 68-84% (w/w) CBGV of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoid CBG. More preferably still the CBGcomprises 19-23% (w/w) of the total phytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid THCA and the phytocannabinoid containingcomponent comprises 54-66% (w/w) of the cannabis plant extract.Preferably the extract further comprises 71-86% (w/w) THCA of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoid THC. More preferably still the THCcomprises 13.4-16.4% (w/w) of the total phytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid CBDA and the phytocannabinoid containingcomponent comprises 71-86% (w/w) of the cannabis plant extract.Preferably the extract further comprises 78-86% (w/w) CBDA of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoid CBD. More preferably still the CBDcomprises 6.1-7.5% (w/w) of the total phytocannabinoid fraction.

In a further embodiment the cannabis plant extract comprises theprinciple phytocannabinoid THCVA and the phytocannabinoid containingcomponent comprises 62-75% (w/w) of the cannabis plant extract.Preferably the extract further comprises 53-65% (w/w) THCVA of the totalphytocannabinoid fraction. More preferably the extract further comprisesthe secondary phytocannabinoid THCV. More preferably still the THCVcomprises 17.3-21.2% (w/w) of the total phytocannabinoid fraction.

In a fourth aspect of the present invention there is provided one ormore phytocannabinoids, either in an isolated form or in the form of abotanical drug substance (BDS), as a prophylactic or in the treatment ofcancer

In a fifth aspect of the present invention there is provided one or morephytocannabinoids taken from the group selected from: THCV, CBDV, THCVA,THCA, CBDA, CBD, CBG, and CBC, for use in the treatment of prostatecancer, wherein the THCVA is present as an isolated phytocannabinoid,the THCA, CBDA CBD, CBG or CBC are present in the form of a BDS, and theTHCV or CBDV are present in either an isolated form or in the form of aBDS.

In accordance with a sixth aspect of the present invention there isprovided the use of one or more phytocannabinoids taken from the groupselected from: THCV, CBDV, THCVA, THCA, CBDA, CBD, CBG, and CBC, for usein the manufacture of a medicament to treat prostate cancer, wherein theTHCVA is present as an isolated phytocannabinoid, the THCA, CBDA CBD,CBG or CBC are present in the form of a BDS, and the THCV or CBDV arepresent in either an isolated form or in the form of a BDS.

In accordance with a seventh aspect of the present invention there isprovided a method of treating a patient with prostate cancer comprisingadministering an effective amount of one or more phytocannabinoids,selected from the group consisting of: THCV, CBDV, THCVA, THCA, CBDA,CBD, CBG, and CBC, wherein, where present, the THCVA is present as anisolated phytocannabinoid, the THCA, CBDA, CBD, CBG or CBC are presentin the form of a BDS, and the THCV or CBDV are present in either anisolated form or in the form of a BDS to the patient.

In one embodiment the one or more phytocannabinoids are propyl variantphytocannabinoids.

In a second embodiment the one or more phytocannabinoids are in an acidform.

In a further embodiment the one or more phytocannabinoids are in aneutral or decarboxylated form.

In a preferred embodiment the phytocannabinoid is CBG and is in the formof a BDS.

Preferably the prostate cancer is hormone-sensitive prostate cancer.

In another embodiment the phytocannabinoid is THCVA in an isolated form.

In a further embodiment the prostate cancer is hormone-insensitiveprostate cancer and the phytocannabinoid is CBD and is in the form of aBDS or the phytocannabinoid is CBDV and is in the form of a BDS.

Preferably the one or more phytocannabinoids are used in combination oras an adjunct therapy with a chemotherapeutic agent and/or ananti-androgen.

Preferably the chemotherapeutic agent is a mitotic inhibitor. Themitotic inhibitor is preferably from the taxane drug class. Morepreferably the mitotic inhibitor taken from the taxane drug class istaken from the group: docetaxel; larotaxel; ortataxel; paclitaxel; andtesetaxel.

When the one or more phytocannabinoids are used in combination with achemotherapeutic agent and or anti-androgen the phytocannabinoid ispreferably CBG or CBD, which may be in the form of a BDS.

In a further embodiment the one or more phytocannabinoids are used forthe purpose of slowing down the growth or reducing the volume of aprostate cancer tumour.

In accordance with a eighth aspect of the present invention there isprovided the use of one or more propyl phytocannabinoids or acidphytocannabinoids for use in the down regulation of ERK signalling andeffect one or more of: anti-proliferation, anti-metastasis oranti-angiogenesis in a human patient.

In accordance with a ninth aspect of the present invention there isprovided the use of one or more propyl phytocannabinoids or acidphytocannabinoids in the manufacture of a medicament to down regulateERK signalling and effect one or more of: anti-proliferation,anti-metastasis or anti-angiogenesis in a human patient.

In accordance with a tenth aspect of the present invention there isprovided a method of treating a patient with cancer comprisingadministering one or more propyl phytocannabinoids or acidphytocannabinoids to down regulate ERK signalling and effect one or moreof: anti-proliferation, anti-metastasis or anti-angiogenesis to thepatient.

Preferably the one or more phytocannabinoids are selected from the groupconsisting of: THCV, CBGV, CBDV, CBGA and CBDA.

Preferably the one or more phytocannabinoids are in an isolated form.

Preferably the one or more propyl or acid phytocannabinoids are for usein the treatment of lung cancer, prostate cancer, or breast cancer.

Preferably the one or more propyl or acid phytocannabinoids are for usein the treatment of bone or lymph metastasis.

In accordance with a eleventh aspect of the present invention there isprovided the use of one or more phytocannabinoid acids, excluding CBDAor CBDVA, for use in medicine.

In accordance with an twelfth aspect of the present invention there isprovided the use of the one or more phytocannabinoid acids for use inthe treatment of cancer.

In accordance with a thirteenth aspect of the present invention there isprovided the use of one or more phytocannabinoid acids in themanufacture of a medicament for use in the treatment of cancer.

In accordance with a fourteenth aspect of the present invention there isprovided a method of treating a patient with cancer comprisingadministering a therapeutic amount of one or more phytocannabinoid acidsto the patient.

Preferably the one or more phytocannabinoid acids are in the form of aBDS.

Preferably the cancer to be treated is a cancer of the prostate, breast,colon, lung, glioma or skin.

Preferably the phytocannabinoid acid is taken from the group consistingof: THCA, CBGA and CBDA.

More preferably there is provided a combination of the phytocannabinoidTHCA with CBDA and/or CBGA.

In accordance with a fifteenth aspect of the present invention there isprovided an isolated CBD, CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDSfor use in the treatment of a pre-cancerous symptom of colon cancer.

In accordance with a sixteenth aspect of the present invention there isprovided the use of an isolated CBD, CBG, CBDV, CBD BDS, CBG BDS and/orCBDV BDS in the manufacture of a medicament to treat a pre-canceroussymptom of colon cancer.

In accordance with a seventeenth aspect of the present invention thereis provided a method of treating a patient with a pre-cancerous symptomof colon cancer, comprising administering a therapeutically effectiveamount of an isolated CBD, CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDSto the patient.

In one embodiment the isolated CBD, CBG, CBDV, CBD BDS, CBG BDS and/orCBDV BDS are used in the treatment of aberrant crypts in the colon.

In a further embodiment the isolated CBD, CBG, CBDV, CBD BDS, CBG BDSand/or CBDV BDS are used in the treatment of colon polyps.

In accordance with a eighteenth aspect of the present invention there isprovided a combination of phytocannabinoids together with achemotherapeutic agent which is not a cannabinoid, for use in thetreatment of a glioma.

In accordance with a nineteenth aspect of the present invention there isprovided the use of a combination of phytocannabinoids together with achemotherapeutic agent which is not a cannabinoid, in the manufacture ofa medicament to treat a glioma.

In accordance with a twentieth aspect of the present invention there isprovided a method of treating a patient with a glioma, comprisingadministering a therapeutically effective amount of a combination ofphytocannabinoids together with a chemotherapeutic agent which is not acannabinoid, to the patient.

Preferably the combination of phytocannabinoids and the chemotherapeuticagent which is not a cannabinoid are packaged for administrationseparately, simultaneously or sequentially.

Preferably the phytocannabinoids are THC and CBD.

Preferably the dose level of the phytocannabinoids is sub-effective forthe treatment of the glioma if used alone.

Preferably the chemotherapeutic agent is temazolamide.

Preferably the dose level of the temazolamide is sub-effective for thetreatment of glioma if used alone.

BRIEF DESCRIPTION OF THE DRAWINGS

Embodiments of the invention are further described hereinafter withreference to the accompanying drawings, in which:

FIG. 1 shows an overview of Botanical Drug Substance (BDS) preparation;

FIG. 2 shows the effect of cannabinoids on apoptosis inhormone-insensitive prostate cancer cell line (DU-145) andhormone-sensitive prostate cancer cell line (LNCaP);

FIG. 3 shows the effect of cannabinoid BDS real and reconstituted onTRPM8 antagonism;

FIG. 4(A-D) shows the effect of cannabinoids alone or in combinationwith a chemotherapeutic agent in hormone-insensitive prostate cancercell line (DU-145) and hormone-sensitive prostate cancer cell line(LNCaP) in a subcutaneous xenograft model;

FIG. 5 (a-c) shows the effect of the cannabinoids isolated CBD and CBDBDS on colon carcinogenesis in the mouse; and

FIG. 6 (a-c) shows the effect of the cannabinoids isolated CBG, isolatedCBDV, CBG BDS and CBDV BDS on colon carcinogenesis in the mouse.

DETAILED DESCRIPTION

The use of phytocannabinoids, either isolated or with all theirco-extracted components is described in the following examples.

EXAMPLE 1 Botanical Production and Manufacture of Cannabinoid BDS

Botanical Raw Material (BRM) is obtained from varieties of Cannabissativa L. (chemotypes) which have been developed to specifically producehigh levels of a given phytocannabinoid as the principalphytocannabinoid. The cannabinoid CBG is the precursor molecule in thebiosynthetic pathway to THC, CBD and CBC. Other cannabinoids are thenformed from these cannabinoids. The principal cannabinoid produced inthe plant will be present as the carboxylic acid form in the plantmaterial, as are any of the other secondary or minor cannabinoids. Thecarboxylic acid form of the cannabinoid is usually decarboxylated to theneutral form during processing of the BRM to Botanical Drug Substance(BDS).

The plants used to prepare the cannabinoid BDS can either be wild typeplants or plants specifically bred to produce one cannabinoid as aprinciple cannabinoid. These plants are referred to as ‘chemotypes’. Forexample the paper by (De Meijer & Hammond, 2005) describes the selectivebreeding of a plant high in CBG. Wild type plants that produce a largeamount of CBG have been found in European fibre hemp populations.

Botanical Drug Substance (BDS) are prepared from BRM and are extractssuitable for further formulation and/or investigative purposes. Theseextracts are termed a Botanical Drug Substance. A brief overview of themethod is provided in FIG. 1. Conditions for the extraction process areoptimised to give the most favourable balance of cannabinoid content andnon-cannabinoid fraction along with satisfactory yield. For example thecannabinoid content of the CBG BDS is virtually 100% CBG, with only verysmall quantities of other cannabinoids present.

Cannabinoid free BDS can be prepared from CBG BDS or a zero cannabinoidplant such as USO-31. Because CBG is the major cannabinoid present inCBG BDS it is possible to remove the CBG present relatively easily usingstandard techniques known in the art such as column chromatography. ACBG-free extract can be used to assess what pharmacology if any, thereis associated with the non-cannabinoid fraction. It is possible tofractionate the BDS completely so that individual compounds can beremoved for purification and the remainder recombined to produce,following solvent removal, a BDS free of the selected compound(s). TheCBG free extract thus produced allows for the evaluation of any synergybetween the cannabinoid and non-cannabinoid fractions.

Isolated phytocannabinoids can also be prepared. As indicated abovecolumn chromatography may be used to isolate CBG from CBG BDS to producepurity greater than 99%. The BDS and the isolated phytocannabinoid canthen be used to compare the effectiveness and any synergy between theprinciple phytocannabinoid and the other phytocannabinoids andnon-cannabinoid constituents in the BDS.

EXAMPLE 2 Phytocannabinoid and Non-Cannabinoid Components in BDS

The following example illustrates the different phytocannabinoidcomponents that make up each of the BDS's described. In each table theprinciple phytocannabinoid is defined in bold typeface.

The BDS's were extracted using liquid CO₂ and then a high performanceliquid chromatography (HPLC) method was used to analyse the differentcannabinoid components in each cannabinoid BDS.

The tables detailed below describe average amounts of the principle,secondary and minor phytocannabinoids in each representative BDS. Theskilled person will appreciate that as the BDS's are extracted fromcannabis plants they will of course be subject to a degree of variationin their composition. Generally the amounts by which each of thephytocannabinoid components will vary by will be in the range of ±10%(w/w). However depending on the starting plant material and the methodof extraction used these amounts may vary by as little as ±5% up to ±50%(w/w).

TABLE 2.1.1 Cannabigerol BDS amount in total and range Amount RangeRange Range CBG BDS (% w/w) (±10%) (±25%) (±50%) CBGV 0.33 0.30-0.360.25-0.41 0.17-0.50 CBG 66.96 60.3-73.7 50.2-83.7  33.5-100.0 THC 0.030.027-0.033 0.023-0.038 0.015-0.045 CBC 0.07 0.06-0.08 0.05-0.090.035-0.105 CBG (related 1.35 1.22-1.49 1.01-1.69 0.68-2.03 substance)Total 68.74 Cannabinoids Total Non- 31.26 cannabinoids

The total phytocannabinoid containing fraction of CBG BDS comprisesapproximately 61-75% (w/w) of the total BDS.

TABLE 2.1.2 Cannabigerol BDS by percentage cannabinoid Amount CBG BDS (%of total cannabinoid) CBGV 0.48 CBG 97.41 THC 0.04 CBC 0.10 CBG (relatedsubstance) 1.96

The amount of the principle phytocannabinoid in the CBG BDS as apercentage of the phytocannabinoid containing fraction is approximately88-100% (w/w).

TABLE 2.2.1 Tetrahydrocannabinol BDS amount in total and range AmountRange Range Range THC BDS (% w/w) (±10%) (±25%) (±50%) CBO 0.2 0.18-0.220.15-0.25 0.1-0.3 CBG 2.0 1.8-2.2 1.5-2.5 1.0-3.0 CBD 1.0 0.9-1.10.75-1.25 0.5-1.5 THCV 1.1 0.99-1.21 0.83-1.38 0.55-1.65 CBN 3.0 2.7-3.32.25-3.75 1.5-4.5 THC (related 0.6 0.54-0.66 0.45-0.75 0.3-0.9substances) THC 74.0 66.6-81.4 55.5-92.5  37.0-100.0 CBC 2.0 1.8-2.21.5-2.5 1.0-3.0 THCA 1.5 1.35-1.65 1.13-1.88 0.75-2.25 Total 85.40Cannabinoids Total Non- 14.60 cannabinoids

The total phytocannabinoid containing fraction of THC BDS comprisesapproximately 77-94% (w/w) of the total BDS.

TABLE 2.2.2 Tetrahydrocannabinol BDS by percentage cannabinoid AmountTHC BDS (% of total cannabinoid) CBO 0.23 CBG 2.34 CBD 1.17 THCV 1.29CBN 3.51 THC (related substances) 0.70 THC 86.65 CBC 2.34 THCA 1.76

The amount of the principle phytocannabinoid in the THC BDS as apercentage of the phytocannabinoid containing fraction is approximately78-95% (w/w).

TABLE 2.3.1 Cannabidiol BDS amount in total and range Amount Range RangeRange CBD BDS (% w/w) (±10%) (±25%) (±50%) CBD (related 0.3 0.27-0.330.23-0.38 0.15-0.45 substances) CBDV 1.9 1.71-2.09 1.43-2.38 0.95-2.85CBDA 1.3 1.17-1.43 0.98-1.63 0.65-1.95 CBG 2.5 2.25-2.75 1.88-3.131.25-3.75 CBN 0.2 0.18-0.22 0.15-0.25 0.1-0.3 CBD 70.0 63.0-77.052.5-87.5  35.0-100.0 THC 5.5 4.95-6.05 4.13-6.88 2.75-8.25 CBC 5.65.04-6.16 4.20-7.00 2.80-8.40 Total 87.30 Cannabinoids Total Non- 12.70cannabinoids

The total phytocannabinoid containing fraction of CBD BDS comprisesapproximately 79-96% (w/w) of the total BDS.

TABLE 2.3.2 Cannabidiol BDS by percentage cannabinoid Amount CBD BDS (%of total cannabinoid) CBD (related substances) 0.34 CBDV 2.18 CBDA 1.49CBG 2.86 CBN 0.23 CBD 80.18 THC 6.30 CBC 6.41

The amount of the principle phytocannabinoid in the CBD BDS as apercentage of the phytocannabinoid containing fraction is approximately72-88% (w/w).

TABLE 2.4.1 Cannabichromene BDS amount in total and range Amount RangeRange Range CBC BDS (% w/w) (±10%) (±25%) (±50%) CBG 0.91 0.82-1.000.68-1.14 0.46-1.37 CBD 3.96 3.56-4.36 2.97-4.95 1.98-5.94 CBCV 0.740.67-0.81 0.56-0.93 0.37-1.11 THC 1.76 1.58-1.94 1.32-2.20 0.88-2.64 CBC(related 0.13 0.12-0.14 0.10-0.16 0.07-0.20 substances) CBC 42.9538.65-47.25 32.22-56.69 21.48-64.43 CBCA 0.56 0.50-0.62 0.42-0.700.28-0.84 CBL 3.54 3.19-3.89 2.67-4.43 1.77-5.31 Total 54.55Cannabinoids Total Non- 45.45 cannabinoids

The total phytocannabinoid containing fraction of CBC BDS comprisesapproximately 49-60% (w/w) of the total BDS.

TABLE 2.4.2 Cannabichromene BDS by percentage cannabinoid Amount CBC BDS(% of total cannabinoid) CBG 1.67 CBD 7.26 CBCV 1.36 THC 3.23 CBC(related substances) 0.24 CBC 78.74 CBCA 1.03 CBL 6.49

The amount of the principle phytocannabinoid in the CBC BDS as apercentage of the phytocannabinoid containing fraction is approximately71-87% (w/w). The CBC BDS also has two secondary cannabinoids: CBD whichis present at approximately 6.5-8% (w/w) of the phytocannabinoidcontaining fraction and CBL which is present at approximately 5.8-7.1%(w/w) of the phytocannabinoid containing fraction.

TABLE 2.5.1 Tetrahydrocannabivarin BDS amount in total and range AmountRange Range Range THCV BDS (% w/w) (±10%) (±25%) (±50%) CBGV 0.150.14-0.17 0.11-0.19 0.07-0.23 CBNV 1.30 1.20-1.40 1.00-1.60 0.65-1.95THCV 64.49 58.04-70.94 48.37-80.61 32.25-96.74 CBCV 0.65 0.59-0.720.49-0.81 0.33-0.98 THC-C4 0.82 0.74-0.90 0.62-1.03 0.41-1.23 CBN 0.150.14-0.17 0.11-0.19 0.07-0.23 THCVA 0.36 0.32-0.40 0.27-0.45 0.18-0.54THC 13.43 12.09-14.77 10.07-16.79  7.72-20.15 Unknowns 0.58 0.52-0.640.44-0.73 0.29-0.87 Total 81.93 Cannabinoids Total Non- 18.07cannabinoids

The total phytocannabinoid containing fraction of THCV BDS comprisesapproximately 74-90% (w/w) of the total BDS.

TABLE 2.5.2 Tetrahydrocannabivarin BDS by percentage cannabinoid AmountTHCV BDS (% of total cannabinoid) CBGV 0.18 CBNV 1.59 THCV 78.71 CBCV0.79 THC-C4 1.00 CBN 0.18 THCVA 0.44 THC 16.39 Unknowns 0.71

The amount of the principle phytocannabinoid in the THCV BDS as apercentage of the phytocannabinoid containing fraction is approximately71-87% (w/w). The THCV BDS also has a secondary cannabinoid THC which ispresent at approximately 14.8-18% (w/w) of the phytocannabinoidcontaining fraction.

TABLE 2.6.1 Cannabidivarin BDS amount in total and range Amount RangeRange Range CBDV BDS (% w/w) (±10%) (±25%) (±50%) CBDVA 0.14 0.13-0.150.11-0.18 0.07-0.21 CBDV 41.19 37.07-45.31 30.89-51.49 20.60-61.79 CBDA0.07 0.06-0.08 0.05-0.09 0.04-0.11 CBG 0.59 0.53-0.65 0.44-0.740.30-0.89 CBD 17.70 15.93-19.47 13.28-22.13  8.85-26.55 THCV 3.062.75-6.12 2.30-3.83 1.53-4.59 CBCV 4.35 3.92-4.79 3.26-5.44 2.18-6.53THC 0.88 0.79-0.97 0.66-1.10 0.44-1.32 CBDV (related 2.20 1.98-2.421.65-2.75 1.10-3.30 substances) CBC 0.93 0.84-1.02 0.70-1.16 0.47-1.40Total 71.11 Cannabinoids Total Non- 28.89 cannabinoids

The total phytocannabinoid containing fraction of CBDV BDS comprisesapproximately 64-78% (w/w) of the total BDS.

TABLE 2.6.2 Cannabidivarin BDS by percentage cannabinoid Amount CBDV BDS(% of total cannabinoid) CBDVA 0.20 CBDV 57.92 CBDA 0.10 CBG 0.83 CBD24.89 THCV 4.30 CBCV 6.12 THC 1.24 CBDV (related substances) 3.09 CBC1.31

The amount of the principle phytocannabinoid in the CBDV BDS as apercentage of the phytocannabinoid containing fraction is approximately52-64% (w/w). The CBDV BDS also has two secondary cannabinoids: CBDwhich is present at approximately 22.4-27.4% (w/w) of thephytocannabinoid containing fraction and CBCV which is present atapproximately 5.5-6.7% (w/w) of the phytocannabinoid containingfraction.

TABLE 2.7.1 Cannabigerol propyl variant BDS amount in total and rangeAmount Range Range Range CBGV BDS (% w/w) (±10%) (±25%) (±50%) CBGV45.92 41.33-50.51 34.44-57.40 22.96-68.88 CBG 12.79 11.51-14.07 9.59-15.99  6.40-19.19 THC 0.08 0.07-0.09 0.06-0.10 0.04-0.12 CBC 0.210.19-0.23 0.16-0.25 0.11-0.32 CBG (related 1.45 1.31-1.60 1.09-1.810.73-2.18 substances) Total 60.45 Cannabinoids Total Non- 39.55cannabinoids

The total phytocannabinoid containing fraction of CBGV BDS comprisesapproximately 54-66% (w/w) of the total BDS.

TABLE 2.7.2 Cannabigerol propyl variant BDS by percentage cannabinoidAmount CBGV BDS (% of total cannabinoid) CBGV 75.96 CBG 21.16 THC 0.13CBC 0.35 CBG (related substances) 2.40

The amount of the principle phytocannabinoid in the CBGV BDS as apercentage of the phytocannabinoid containing fraction is approximately68-84% (w/w). The CBGV BDS also has a secondary cannabinoid CBG which ispresent at approximately 19-23% (w/w) of the phytocannabinoid containingfraction.

TABLE 2.8.1 Tetrahydrocannabinolic acid BDS amount in total and rangeAmount Range Range Range THCA BDS (% w/w) (±10%) (±25%) (±50%) CBO 0.060.05-0.07 0.05-0.08 0.03-0.09 CBG 1.91 1.72-2.10 1.43-2.39 0.96-2.87 CBD0.30 0.27-0.33 0.23-0.38 0.15-0.45 THC (related 0.16 0.14-0.18 0.12-0.200.08-0.24 substances) THCV 0.05 0.04-0.06 0.04-0.06 0.03-0.08 CBN 1.111.00-1.22 0.83-1.39 0.56-1.67 THC 8.93 8.04-9.82  6.70-11.16  4.47-13.40CBL 0.17 0.15-0.19 0.13-0.21 0.09-0.26 CBC 0.26 0.23-0.29 0.20-0.330.13-0.39 THCA 46.98 42.28-51.68 35.24-58.73 23.49-70.47 Total 59.93Cannabinoids Total Non- 40.07 cannabinoids

The total phytocannabinoid containing fraction of THCA BDS comprisesapproximately 54-66% (w/w) of the total BDS.

TABLE 2.8.2 Tetrahydrocannabinolic acid BDS by percentage cannabinoidAmount THCA BDS (% of total cannabinoid) CBO 0.10 CBG 3.19 CBD 0.50 THC(related substances) 0.27 THCV 0.08 CBN 1.85 THC 14.90 CBL 0.28 CBC 0.43THCA 78.39

The amount of the principle phytocannabinoid in the THCA BDS as apercentage of the phytocannabinoid containing fraction is approximately71-86% (w/w). The THCA BDS also has a secondary cannabinoid THC which ispresent at approximately 13.4-16.4% (w/w) of the phytocannabinoidcontaining fraction.

TABLE 2.9.1 Cannabidiolic acid BDS amount in total and range AmountRange Range Range CBDA BDS (% w/w) (±10%) (±25%) (±50%) CBDV 0.230.21-0.25 0.17-0.29 0.12-0.37 CBDA 68.14 61.33-74.95 51.11-85.1834.07-100.0 CBD 5.36 4.82-5.90 4.02-6.70 2.68-8.04 CBN 0.19 0.17-0.210.14-0.24  0.1-0.29 THC 0.53 0.48-0.58 0.40-0.66 0.27-0.80 CBL 0.290.26-0.32 0.22-0.36 0.15-0.44 CBC 0.38 0.34-0.42 0.29-0.48 0.19-0.57 CBD(related 3.31 2.98-3.64 2.48-4.14 1.66-4.98 substances) Total 78.43Cannabinoids Total Non- 21.57 cannabinoids

The total phytocannabinoid containing fraction of CBDA BDS comprisesapproximately 71-86% (w/w) of the total BDS.

TABLE 2.9.2 Cannabidiolic acid BDS by percentage cannabinoid Amount CBDABDS (% of total cannabinoid) CBDV 0.29 CBDA 86.88 CBD 6.83 CBN 0.24 THC0.68 CBL 0.37 CBC 0.48 CBD (related substances) 4.22

The amount of the principle phytocannabinoid in the CBDA BDS as apercentage of the phytocannabinoid containing fraction is approximately78-96% (w/w). The CBDA BDS also has a secondary cannabinoid CBD which ispresent at approximately 6.1-7.5% (w/w) of the phytocannabinoidcontaining fraction.

TABLE 2.10.1 Tetrahydrocannabivarinic acid BDS amount in total and rangeAmount Range Range Range THCVA BDS (% w/w) (±10%) (±25%) (±50%) CBDVA1.44 1.30-1.58 1.08-1.80 0.72-2.16 CBNV 0.35 0.32-0.39 0.26-0.440.18-0.53 THCV 13.17 11.85-14.49  9.88-16.46  6.59-19.76 CBCV 1.971.77-2.17 1.48-2.46 0.99-2.96 THC-C4 0.36 0.32-0.40 0.27-0.45 0.18-0.54THCVA 40.61 36.55-44.67 30.46-50.76 20.31-50.76 THC 3.53 3.18-3.882.65-4.41 1.77-5.30 CBC 0.20 0.18-0.22 0.15-0.25 0.1-0.3 THCA 6.295.66-6.92 4.72-7.86 3.15-9.44 Unknowns 0.45 0.41-0.50 0.38-0.560.23-0.68 Total 68.37 Cannabinoids Total Non- 31.63 cannabinoids

The total phytocannabinoid containing fraction of THCVA BDS comprisesapproximately 37-45% (w/w) of the total BDS.

TABLE 2.10.2 Tetrahydrocannabivarinic acid BDS by percentage cannabinoidAmount THCVA BDS (% of total cannabinoid) CBDVA 2.11 CBNV 0.51 THCV19.26 CBCV 2.88 THC-C4 0.53 THCVA 59.40 THC 4.90 CBC 0.29 THCA 9.20Unknowns 0.66

The amount of the principle phytocannabinoid in the THCVA BDS as apercentage of the phytocannabinoid containing fraction is approximately53-65% (w/w). The THCVA BDS also a secondary cannabinoid THCV which ispresent at approximately 17.3-21.2% (w/w) of the phytocannabinoidcontaining fraction.

The following table details an overview of the amount of cannabinoid andnon-cannabinoid fraction in each cannabinoid BDS and the amount ofcannabinoid as a percentage of the total cannabinoids in each BDS. Aspreviously discussed the skilled person will appreciate that thesevalues will vary due to the naturally occurring nature of the startingplant material.

TABLE 2.11.1 Overview of cannabinoid BDS Cannabinoid Amount of principleFraction Non-cannabinoid cannabinoid (% of BDS (% w/w) Fraction (% w/w)total cannabinoid) CBG 68.7 31.3 97.4 THC 85.4 14.6 86.7 CBD 87.3 12.780.2 CBC 54.5 45.5 78.7 THCV 81.9 18.1 78.7 CBDV 71.1 28.9 57.9 CBGV60.5 39.5 76.0 THCA 59.9 40.1 78.4 CBDA 78.4 21.6 86.9 THCVA 68.4 31.659.4

It is desirable to maximise the amount of principle phytocannabinoid inthe phytocannabinoid containing fraction, however in some cases synergymay exist between the principle and secondary cannabinoids which maylead to enhanced medicinal effects.

It is also desirable for the range by which the percentage of thephytocannabinoid containing fraction, the non-phytocannabinoidcontaining fraction and the amount of principle phytocannabinoid varies.In most cases this variation will be small and be in the range of ±5%,up to ±10%, up to ±25% and preferably no greater than ±50%.

The non-cannabinoid components of a phytocannabinoid BDS may play animportant role in the BDS's pharmacology. As such the terpene profile isclassified below. The following tables illustrate the terpene profile ofa CBD chemovar which is representative of a high phytocannabinoidcontaining plant. Five plants were freshly harvested and extracted usingsteam distillation. The principle monoterpene and sesquiterpene arehighlighted in bold.

TABLE 2.12.1 Monoterpene amount by percentage of total terpene fractionand ranges Amount (% of terpene Range Range Range Monoterpenes fraction)(±10%) (±25%) (±50%) Pinene (alpha 10.56  9.50-11.62  7.92-13.20 5.28-15.84 & beta) Myrcene 39.46 35.51-43.41 29.60-49.33 19.73-59.19Limonene 4.14 3.73-4.55 3.11-5.18 2.07-6.21 Beta-ocimene 4.04 3.64-4.443.03-5.05 2.02-6.06 Total 58.20

The monoterpene containing fraction comprises approximately 52-64% (w/w)of the total terpene fraction.

TABLE 2.12.2 Monoterpene amount by percentage of monoterpenes AmountMonoterpenes (% of monoterpene fraction) Pinene (alpha & beta) 18.14Myrcene 67.80 Limonene 7.12 Beta-ocimene 6.94

The amount of the principle monoterpene myrcene in the monoterpenefraction as a percentage of the monoterpene fraction is approximately61-75% (w/w). The monoterpene fraction also has a secondary monoterpenepinene which is present at approximately 16.3-20% (w/w) of themonoterpene fraction.

TABLE 2.12.3 Sesquiterpene amount by percentage of total terpenefraction and ranges Amount (% of terpene Range Range RangeSesquiterpenes fraction) (±10%) (±25%) (±50%) Caryophyllenes 29.2726.34-32.20 21.95-36.59 14.64-43.91 (t & oxide) Bergotamene 0.180.16-0.20 0.14-0.23 0.09-0.27 Humulene 7.97 7.17-8.77 5.98-9.96 3.99-11.96 Aromaden- 0.33 0.30-0.36 0.25-0.41 0.17-0.50 drene Selinene0.59 0.53-0.65 0.44-0.74 0.30-0.89 Anon 0.44 0.40-0.48 0.33-0.550.22-0.66 Farnesene 1.55 1.40-1.71 1.16-1.94 0.78-2.33 (Z, E & alpha)alpha 0.12 0.11-0.13 0.09-0.15 0.06-0.18 Gurjunene Bisabolene 0.390.35-0.43 0.29-0.49 0.20-0.59 Nerolidol 0.43 0.39-0.47 0.32-0.540.22-0.65 Diepicedrene- 0.38 0.34-0.42 0.29-0.48 0.19-0.57 1-oxideAlpha- 0.16 0.14-0.18 0.12-0.20 0.08-0.24 Bisabolol Total 41.80

The sesquiterpene containing fraction comprises approximately 27-32%(w/w) of the total terpene fraction.

TABLE 2.12.4 Sesquiterpene amount by percentage of sesquiterpenes AmountSesquiterpenes (% of sesquiterpene fraction) Caryophyllenes (t & oxide)70.02 Bergotamene 0.43 Humulene 19.07 Aromadendrene 0.79 Selinene 1.41Anon 1.05 Farnesene (Z, E & alpha) 3.71 alpha Gurjunene 0.29 Bisabolene0.93 Nerolidol 1.03 Diepicedrene-1-oxide 0.91 Alpha-Bisabolol 0.38

Patent application number PCT/GB2008/001837 describes the production ofa ‘zero cannabinoid’ plant. These plants were produced by selectivebreeding to produce a Cannabis sativa L plant that contained a generallyqualitatively similar terpene profile as a Cannabis sativa L plant thatproduced cannabinoids yet it was devoid of any cannabinoids. Theseplants can be used to produce cannabinoid-free plant extracts which areuseful control plants in experiments and clinical trials. A breakdown ofthe terpene profile produced in the plants can be found in the tablebelow. The primary monoterpenes and sesquiterpene are highlighted inbold.

TABLE 2.13.1 Monoterpene amount by percentage of total terpene fractionand ranges Amount (% of terpene Range Range Range Monoterpenes fraction)(±10%) (±25%) (±50%) Pinene 29.34 26.41-32.27 22.01-36.68 14.67-44.01(alpha & beta) Myrcene 29.26 26.33-32.19 21.95-36.58 14.63-43.89Limonene 5.32 4.79-5.85 3.99-6.65 2.66-7.98 Linalol 4.50 4.05-4.953.38-5.63 2.25-6.75 Verbenol 3.45 3.11-3.80 2.59-4.31 1.73-5.18 (cis &trans) Total 71.87

The monoterpene containing fraction comprises approximately 65-79% (w/w)of the total terpene fraction.

TABLE 2.13.2 Monoterpene amount by percentage of monoterpenes Amount (%of monoterpene Monoterpenes fraction) Pinene (alpha & beta) 40.82Myrcene 40.71 Limonene 7.41 Linalol 6.26

The zero cannabinoid plant was found to comprise two principlemonoterpenes; pinene and myrcene. The amount of the principlemonoterpene myrcene in the monoterpene fraction as a percentage of themonoterpene fraction is approximately 37-45% (w/w). The amount of theprinciple monoterpene pinene in the monoterpene fraction as a percentageof the monoterpene fraction is approximately 37-45% (w/w).

TABLE 2.13.3 Sesquiterpene amount by percentage of total terpenefraction and ranges Amount (% of terpene Range Range RangeSesquiterpenes fraction) (±10%) (±25%) (±50%) Caryophyllenes 10.89 9.80-11.98  8.17-13.61  5.45-16.34 (t & oxide) Bergotamene 2.512.26-2.76 1.88-3.14 1.26-3.77 Farnesene (Z, 3.43 3.09-3.77 2.57-4.291.72-5.15 E & alpha) Humulene (& 5.04 4.54-5.54 3.78-6.30 2.52-7.56epoxide II) delta guaiene 2.40 2.16-2.64 1.80-3.00 1.20-3.60 Bisabolene3.85 3.47-4.24 2.89-4.81 1.93-5.78 Total 28.12

The sesquiterpene containing fraction comprises approximately 25-31%(w/w) of the total terpene fraction.

TABLE 2.12.4 Sesquiterpene amount by percentage of sesquiterpenes Amount(% of sesquiterpene Sesquiterpenes fraction) Caryophyllenes (t & oxide)38.73 Bergotamene 8.93 Farnesene (Z, E & alpha) 12.20 Humulene (&epoxide II) 17.92 delta guaiene 8.53 Bisabolene 13.69

The amount of the principle sesquiterpene caryophylene in thesesquiterpene fraction as a percentage of the sesquiterpene fraction isapproximately 35-43% (w/w). The sesquiterpene fraction also has asecondary sesquiterpene humulene which is present at approximately16-20% (w/w) of the sesquiterpene fraction.

EXAMPLE 3 Effect of Phytocannabinoids on Apoptosis inHormone-insensitive Prostate Cancer Cell Line (DU-145) andHormone-sensitive Prostate Cancer Cell Line (LNCaP)

The effect of CBDV BDS, CBD BDS, THCVA BDS, THCV BDS, isolated THCV andisolated THCVA on the apoptosis of two prostate cancer cell lines weretested using a chemioluminescence assay for caspase 3/7 release. Twodifferent concentrations of cannabinoids were tested; a low dose (10 μM)and a high dose (25 μM). The chemioluminescence that were recorded foreach phytocannabinoid are detailed in FIG. 2.

In the hormone-insensitive prostate cancer cell line (DU-145) thehighest chemioluminescence and as such the highest pro-apoptotic effectswere found with the low dose CBDV BDS and the high dose isolated THCVApurified cannabinoid at 25 μM. The high dose THCV BDS, CBD BDS andisolated THCV also showed apotosis levels over that of the control.

In the hormone-sensitive prostate cancer cell line (LNCaP) the highestchemioluminescence and as such the highest pro-apoptotic effect wasfound with the high dose isolated THCVA. The low and high dose CBDV BDSwere also shown to have apoptotic effects on the cancer cells.

These data infer that both the phytocannabinoid BDS's and isolatedphytocannabinoid may be useful in the treatment of cancer as they areable to exert an apoptotic effect on cancer cells.

EXAMPLE 4 Effect of Phytocannabinoids on the G-protein Coupled ReceptorGPR55

An ERK Alphascreen assay was undertaken using GPR55-HEK cells. Thedifferent cannabinoids tested displayed different pharmacologicalprofiles at GPR55 as is illustrated in Table 4.1 below.

TABLE 4.1 Effect of phytocannabinoids at GPR55 Compound Effect at GPR55CBD (control) Antagonist THCV Antagonist (at low concentrations) CBGAInverse agonist CBGV Antagonist CBDA Antagonist CBDV Antagonist

A significant reduction in the ERK signalling pathway has been shown tolead to induction of apotosis (Chang et al., 2003).

These data suggest that GPR55 expression by cancer cells can beantagonised by all the phytocannabinoids tested making them good targetcompounds for use in the treatment of cancer.

EXAMPLE 5 Effect of Phytocannabinoids on Inhibition of MonoacylglycerideLipase (MAGL) and Diacylglyceride Lipase (DAGL)

The effects of the phytocannabinoid BDS THCA, CBN, CBGA, CBDA and CBCVwere tested to determine whether they were able to inhibit the MAGL andDAGL enzymes which are able to hydrolyse the endogenous cannabinoid 2-AGto arachidonic acid and glycerol. The results generated from theseexperiments are detailed in Table 5.1 below.

TABLE 5.1 Effect of phytocannabinoids on inhibition of monoacylglyceridelipase (MAGL) and diacylglyceride lipase (DAGL) Max Max MAGconcentration DAGL concentration Sample IC₅₀ tested (% inhibition) IC₅₀tested (% inhibition) THCA  46 μM 100 μM  25 μM 50 μM (81.9%) (79.3%)CBN >50 μM 50 μM >50 μM  50 μM (31.5%) (26.9%) CBGA >50 μM 50 μM 30 μM100 μM  (17.2%) (67.5%) CBDA >50 μM 50 μM 23 μM 100 μM  (18.5%) (91.4%)CBCV >50 μM 50 μM >50 μM  50 μM  (4.8%) (18.9%)

As can be seen the phytocannabinoid THCA was most effective atinhibiting DAGL and the all the phytocannabinoid acids (THCA, CBDA andCBGA) were effective at inhibiting MAGL. These data infer that thesephytocannabinoid might be useful in the treatment of cancer as they areable to prevent the endogenous cannabinoid 2-AG from being hydrolysedand as such may prevent cancerous cell formation.

An additional experiment was undertaken to determine whetherphytocannabinoid BDS were more effective at inhibition of MAGL thantheir respective purified counterparts. For this experiment CBG BDS wasused as this BDS was the easiest to purify and reconstitute. Fourdifferent test articles were used: CBG BDS, isolated CBG, CBG-free CBGBDS and CBG reconstituted BDS, whereby the CBG-free BDS (as prepared inExample 1) was spiked with isolated CBG at the same concentration of CBGas CBG BDS. Table 5.2 below details the data obtained.

TABLE 5.2 Effect of BDS, purified compounds and reconstituted BDS oninhibition of monoacylglyceride lipase (MAGL) Max tested on SAMPLE EC50on MAGL MAGL % inhibition Isolated CBG 195.2 μM 100 μM (31.49%) CBG BDS64.33 μM 100 μM (57.93%) CBG-Free BDS 187.0 μM 100 μM (36.55%)Reconstituted CBG BDS 70.28 μM 100 μM (59.31%)

As can be seen in Table 5.2 above the CBG BDS is of a similar potency asthe reconstituted BDS. These data demonstrate a synergistic effectbetween the principle phytocannabinoid and the rest of the cannabinoidand non-cannabinoid components in the BDS.

These data infer that the use of phytocannabinoid BDS's comprising anon-cannabinoid component would be more efficacious in the treatment ofcancer than isolated phytocannabinoids.

EXAMPLE 6 Effect of Phytocannabinoid BDS and ReconstitutedPhytocannabinoid BDS on TRPM8 Antagonism

The CBG BDS and the reconstituted CBG BDS as described in Example 5above were used to determine their effectiveness on TRPM8 antagonism.Also tested for completeness were CBG-free BDS and purified CBG. Thepercentage response against 0.25 μM icilin were observed and are asdetailed in FIG. 3.

As can be seen both CBG BDS and reconstituted CBG BDS gave a much higherresponse rate than either isolated CBG or the CBG-free BDS. These dataagain demonstrate the synergy between the principle phytocannabinoid andthe other components in the BDS.

These data surmise that the use of a phytocannabinoid BDS would be amore effective treatment option in cancer.

EXAMPLE 7 Effect of Phytocannabinoids in Hormone-insensitive ProstateCancer Cell Line (DU-145) and Hormone-sensitive Prostate Cancer CellLine (LNCaP) on Cell Vitality (MTT Assay)

Cells were seeded in presence of FBS in 6 wells multiwell with a densityof 8×10⁴ cells/well (DU-145) or 1×10⁵ cells/well (LNCaP). After 7 hours,cells were starved of dihydrotestosterone (DHT) for 15 hours and treatedwith increased concentrations of compounds for 24 or 48 hours (absenceof serum was maintained during the treatments). Cell viability wasassessed by MTT staining.

Phytocannabinoid BDS and their respective isolated phytocannabinoidswere tested in both the hormone-insensitive (DU-145) prostate cancercells and hormone-sensitive (LNCaP) prostate cancer cells. Threedifferent phytocannabinoids were chosen for this experiment: CBG BDS andisolated compound, CBC BDS and isolated compound and CBD BDS andisolated compound.

Table 7.1 details the data observed in the starved cells. As can be seenin each case the BDS has much more of an effect on the vitality of theprostate cancer cells than its respective isolated phytocannabinoid.

TABLE 7.1 Effect of phytocannabinoid BDS on cell vitality (MTT Assay) -cells starved of dihydrotestosterone (DHT) DU-145 LNCaP IC₅₀ on cellvitality IC₅₀ on cell vitality Sample 24 h 48 h 24 h 48 h CBG 11.8 μM 5.8 μM 10.8 μM  8.1 μM CBG BDS 5.3 μM 4.8 μM 7.0 μM 4.7 μM CBC 7.5 μM5.6 μM 10.8 μM  6.0 μM CBC BDS 4.9 μM 4.5 μM 6.3 μM 5.3 μM CBD 5.5 μM4.9 μM 5.3 μM 5.6 μM CBD BDS 5.0 μM 3.6 μM 5.5 μM 4.3 μM

The experiment was repeated in the hormone-insensitive (DU-145) prostatecancer cells and hormone-sensitive (LNCaP) prostate cancer cells thathad not been starved of DHT. Here just the CBG BDS and isolated CBG areillustrated in Table 7.2 below.

TABLE 7.2 Effect of CBG & CBG BDS on cell vitality (MTT Assay) -non-DHT-starved Cells Sample IC₅₀ on cell vitality LNCaP CBG >25 μMCBG-BDS 21.2 μM DU-145 CBG >25 μM CBG-BDS 24 μm

As is demonstrated in Table 7.2 above the presence of DHT in thenon-starved cells dramatically alters the effectiveness of thephytocannabinoid BDS and the isolated phytocannabinoids.

The data described above again illustrates that the phytocannabinoid BDSare more efficacious and as such a better compound to use in thetreatment of cancer.

EXAMPLE 8 Effect of Phytocannabinoids Alone or in Combination With aChemotherapeutic Agent or an Anti-androgen in Hormone-insensitiveProstate Cancer Cell Line (DU-145) and Hormone-sensitive Prostate CancerCell Line (LNCaP) in a Subcutaneous Xenograft Model

The phytocannabinoids CBG BDS and CBD BDS were used in this experimentto demonstrate the in vivo effectiveness of the phytocannabinoid BDS's.

Cancer cells were maintained in vitro in an appropriate culture mediumand were harvested, washed in culture medium and re-suspended inmatrigel for ready for in vivo administration. 1-2×10⁷ cells in 200 μlwere injected subcutaneously into the left flank of mice. The mice wereanaesthetised and a 0.5 mg 5-alpha-dihydrotestosterone pellet (21-dayrelease) implanted subcutaneously into the scruff of each mouse toencourage tumour growth, and the wound was closed.

Once the implanted tumour volume reached between 100-200 mm (2-3 weeks)mice were allocated to their treatment groups. Treatment commenced onDay 17. The phytocannabinoid BDS was administered i.p. once daily atdoses of 1, 10 & 100 mg/kg cannabinoid BDS for a further 25 days.

In a further experiment the chemotherapeutic agent taxotere (5 mg/kg)i.v. was administered weekly, alone or in combination with 100 mg/kgphytocannabinoid BDS.

The chemotherapeutic agent taxotere was used to exemplify the taxanedrug class which are mitotic inhibitors.

In a further experiment CBD BDS at 100 mg/kg of CBD was evaluated forits action in combination with either 25 or 50 mg/kg of theanti-androgen bicalutamide.

The anti-androgen bicalutamide is an example of a hormone therapy drugused in the treatment of prostate cancer. Hormonal therapies interferewith the production of particular hormones by the body. Prostate cancer(when it is in the hormone sensitive stage) requires the male hormonetestosterone (androgen) in order to grow. The prostate cancer cells havereceptors on their surface which when the hormone attaches to it allowsthe cancer cell to grow.

The anti-androgen bicalutamide is structurally similar to testosteroneand prevents the testosterone from binding to the cancer cell. Withoutthe testosterone the cancer cells grow more slowly or may stop growingcompletely and as such the tumour may shrink as a result.

Mice were evaluated daily by an experienced technician for 4-5 weeks.Tumour dimensions were recorded at day 7 (calliper measurement of lengthand width and tumour cross-sectional area and volume calculated) andrecorded three times weekly and body weight measured weekly.

The human dose equivalent (HED) can be estimated using the followingformula:

${HED} = {{Animal}\mspace{14mu}{dose}\mspace{14mu}\left( {{mg}\text{/}{kg}} \right)\mspace{14mu}{multiplied}\mspace{14mu}{by}\mspace{14mu}\frac{{Animal}\mspace{14mu} K_{m}}{{Human}\mspace{14mu} K_{m}}}$The K_(m) for a mouse is 3 and the K_(m) for a human is 37.

FIGS. 4 (a-f) detail the mean tumour volume and weight recorded in theseexperiments.

FIG. 4 a) shows that the growth rate of the tumour was inhibited in adose dependant manner by CBG BDS. Both the 10 and 100 mg/kg doses of BDSwere significant when compared to the vehicle group. In additiontaxotere significantly inhibited both growth rate and terminal tumourvolume, with a slight synergistic effect detected in the group treatedwith both CBG BDS and taxotere.

FIG. 4 b) shows that the growth rate of the tumour was inhibited in adose dependant manner by CBD BDS. Both the 10 and 100 mg/kg doses of BDSwere significant when compared to the vehicle group. Similarly to theexperiments with CBG BDS the taxotere significantly inhibited bothgrowth rate and terminal tumour volume, with a slight synergistic effectdetected in the group treated with both CBD BDS and taxotere.

Figures c) and d) infer that there is little inhibition of tumour volumein the hormone-insensitive (DU-145) prostate cancer cells.

The data shown in FIGS. 4 e) and f) detail the tumour volume and weightof the cells treated with CBD BDS in combination with the anti-androgenbicalutamide. These data demonstrate that the combination of the two,particularly at the lower dose level of bicalutamide is able to produceboth lower tumour volumes and weights than either the bicalutamide orthe CBD BDS alone.

These data all show that phytocannabinoid BDS are very potent compoundsfor use in the treatment of cancer, particularly when these are combinedwith another compound such as an anti-androgen or a chemotherapeuticagent.

EXAMPLE 9 Effect of Phytocannabinoids on Cell Vitality in Breast CancerCells (Alamar Blue Assay)

Cells were seeded in presence of FBS in 6 wells multiwell and treatedwith increased concentrations of compounds for 72 hours. Cell viabilitywas assessed by Alamar blue staining.

Isolated phytocannabinoids were tested on the MDA-MB-231 breast cancercell line in order to determine whether phytocannabinoids were effectivein other types of cancer other than prostate cancer. The differentisolated phytocannabinoids chosen for this experiment were: CBGA, CBDA,CBDV, CBD, THCV and THC.

Table 9.1 below details the effectiveness of the cannabinoids at killingthe breast cancer cells.

TABLE 9.1 Effect of phytocannabinoids on cell viability of breast cancercells. Compound IC50 (μM) CBGA 2.2 CBDA 3.9 CBDV 5.0 CBD 5.0 THCV 6.5THC >10.0

The phytocannabinoid acids CBGA and CBDA both appear to be the mostefficacious in killing the breast cancer cells and as such make goodtargets for the use of these compounds in the treatment of cancer. Thesedata show that surprisingly the phytocannabinoid acids are more than 50%better than the free phytocannabinoids.

EXAMPLE 10 Effect of Phytocannabinoids on Colon Carcinogenesis in theMouse

The phytocannabinoids isolated CBD, CBG and CBDV and BDS's correspondingto CBD, CBG and CBDV were evaluated in their effect to prevent and treatcolon cancer in the mouse. Aberrant crypt focus (ACF), polyps andtumours were induced in the mouse by the carcinogenic substanceazoxymethane. The phytocannabinoids were given (i.p.) to the mice threetimes per week for a period of three months. The COX-2 inhibitorcelecoxib was used as a positive control.

FIG. 5 (a-c) detail the results obtained from the experiments withisolated CBD and CBD BDS. In FIG. 5 a) it can be seen that the CBD BDSis able to statistically significantly reduce the numbers of ACF permouse in comparison to the control.

FIG. 5 b) shows that the CBD BDS is more effective at reducing thenumber of polyps per mouse at a statistically significant level incomparison to the control animals.

FIG. 5 c) shows that the CBD purified compound significantly reduced thenumbers of tumours per animal.

FIG. 6 (a-c) demonstrate the data obtained for isolated CBG and CBDV inaddition to that obtained for CBG BDS and CBDV BDS.

All of the phyto cannabinoids exerted a protective effect against theexperimentally induced colon carcinogenesis as is shown in FIG. 6 a)with isolated CBG giving the most statistically significant results.

FIG. 6 b) demonstrates that the protective effect was even morepronounced on the formation of ACF with 4 or more crypts. These type ofcrypts are predictive of the final incidence of colon cancer and on theformation of tumours. As is shown isolated CBG gave the moststatistically significant data demonstrating such a protective effectthat there were no ACF with greater than 4 crypts produced in theanimals given isolated CBG.

FIG. 6 c) details the number of tumours that occurred in each animal.Again the data produced in the mice given isolated CBG was such that notumours were produced in these animals.

In summary these data demonstrate the protective effects ofphytocannabinoids in the prevention of colon cancer. Of significance isthe phytocannabinoid CBG which exerts a strong protective effect againstcolon cancer particularly when it is in an isolated form.

EXAMPLE 11 Effect of Phytocannabinoids in Human Glioma Cells on CellVitality (MTT Assay)

Cells were seeded in presence of FBS in 6 wells multiwell with a densityof 8×10⁴ cells/well. Glioma cells were treated with increasedconcentrations of compounds and cell viability was assessed by MTTstaining.

CBD BDS and isolated CBD, CBG and CBDV were tested.

Table 11.1 details the data observed. As can be seen the CBD BDS gavethe best result with the lowest concentration being a little moreeffective than its respective isolated phytocannabinoid.

TABLE 11.1 Effect of phytocannabinoids on cell vitality (MTT Assay)Sample IC₅₀ on glioma cell vitality CBD 8.92 μM CBD BDS 8.83 μM CBG12.40 μM  CBDV 12.40 μM 

As can be observed these data indicate that both CBD BDS and isolatedCBD might be useful treatments in brain tumours including glioma.

EXAMPLE 12 Effect of the Cannabinoids THC and CBD Alone and inCombination With Each Other and/or the Chemotherapeutic AgentTemazolamide on the Viability of Human Glioma Cell Lines (MTT Assay)

The synergistic action of the combined administration of thephytocannabinoids THC and CBD in equal amounts was tested in humanglioma cell lines. The combined administration of the twophytocannabinoids led to a synergic reduction in the viability of humanglioma cells during an MTT assay.

Table 12.1 below shows that the sub-effective dose levels of THC and CBD(0.7 μM) leads to a statistically significant reduction in cellviability when the two cannabinoids are combined. The cell viability isdramatically reduced when the dose level of the THC and CBD areincreased.

TABLE 12.1 Cell viability of U87 glioma cells treated with THC, CBD andTHC:CBD (1:1) Dose level Compound (μM) Cell Viability (%) Control — 100THC 0.7 100 CBD 0.7 115 THC:CBD (1:1) 0.7 (each) 90 THC 0.9 92 CBD 0.995 THC:CBD (1:1) 0.9 (each) 70 THC 1.2 80 CBD 1.2 70 THC:CBD (1:1) 1.2(each) 35

Further experiments were undertaken using the chemotherapeutic agenttemazolamide (TMZ) these data are shown in Table 12.2 below.

TABLE 12.2 Cell viability of U87 glioma cells treated with THC, CBD,THC:CBD (1:1) and TMZ Compound Cell Viability (%) Control 100 THC (1 μM)115 CBD(1 μM) 98 TMZ (100 μM) 82 THC:CBD (1 μM each) 55 THC (1 μM) + TMZ(100 μM) 70 CBD (1 μM) + TMZ (100 μM) 65 THC:CBD (1 μM each) + TMZ (100μM) 38

The combined administration of sub-maximal doses of THC, CBD and TMZ ledto a synergic reduction of the viability of the U87 glioma cells.

EXAMPLE 13 Effect of the Cannabinoids THC and CBD Alone and inCombination With Each Other and/or the Chemotherapeutic AgentTemazolamide on the Viability of Human Glioma Xenografts

Table 13.1 below details the data recorded when the phytocannabinoidsTHC and CBD were tested alone or in combination on the human gliomaxenograft tumours.

TABLE 13.1 Tumour volume after treatment with THC, CBD and THC:CBD (1:1)Dose level Tumour volume Compound (mg/kg) (increase from day 1) Control— 10.3 THC 3.7 8.7 CBD 3.7 9.1 THC:CBD (1:1) 3.7 (each) 8.9 THC 7.5 8.5CBD 7.5 8.9 THC:CBD (1:1) 7.5 (each) 5.7 THC 15.0  5.8 CBD 15.0  7.6

The phytocannabinoids were also tested with the TMZ as is detailed inTable 13.2 below.

TABLE 13.2 Tumour volume after treatment with THC, CBD, THC:CBD (1:1)and TMZ Tumour volume Compound (increase from day 1) Control 10.3THC:CBD (3.7 mg/kg each) 8.9 TMZ (100 μM) 4.1 THC:CBD (3.7 mg/kg each) +TMZ 2.6 (100 μM)The following paragraphs are not claims, but represent preferred aspectsand embodiments of the invention.

1. A cannabis plant extract comprising a phytocannabinoid containingcomponent and a non-phytocannabinoid containing component, for use inmedicine, wherein the phytocannabinoid containing component comprises atleast 50% (w/w) of the cannabis plant extract and thenon-phytocannabinoid containing component comprises a monoterpenefraction and a sesquiterpene fraction, in which a principle monoterpenesub-fraction is selected from myrcenes or pinenes and a principlesesquiterpene sub-fraction is selected from caryophyllenes or humulenes.

2. The use of a cannabis plant extract comprising a phytocannabinoidcontaining component and a non-phytocannabinoid containing component,for use in the manufacture of a medicament for use in medicine, whereinthe phytocannabinoid containing component comprises at least 50% (w/w)of the cannabis plant extract and the non-phytocannabinoid containingcomponent comprises a monoterpene fraction and a sesquiterpene fractionand wherein a principle monoterpene sub-fraction is selected frommyrcenes or pinenes and a principle sesquiterpene sub-fraction isselected from caryophyllenes or humulenes.

3. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraphs 1 and 2, for use in the treatment of cancer.

4. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraphs 1-3, wherein the principle monoterpenesub-fraction comprises myrcenes and the secondary monoterpenesub-fraction comprises pinenes.

5. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraphs 1-4, wherein the principle monoterpenessub-fraction comprises both myrcenes and pinenes.

6. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of the preceding paragraphs, wherein the principlesesquiterpene sub-fraction comprises caryophyllenes and secondarysesquiterpene sub-fraction comprises humulenes.

7. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of the preceding paragraphs, wherein the principlephytocannabinoid is selected from the group consisting of: THCV, CBDV,CBGV, THCVA, THCA, CBDA, CBG, THC, CBD and CBC.

8. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of the preceding paragraphs, wherein thenon-phytocannabinoid containing component further comprises one or morecompounds from the group consisting of: diterpenes; triterpenes;sterols; triglycerides; alkanes; squalenes; tocopherols; andcarotenoids.

9. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of the preceding paragraphs, wherein the principlephytocannabinoid is CBG and the phytocannabinoid containing componentcomprises 61-75% (w/w) of the cannabis plant extract.

10. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 9, comprising greater than 88% (w/w) CBG of thetotal phytocannabinoid fraction.

11. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis THC and the phytocannabinoid containing component comprises 77-94%(w/w) of the cannabis plant extract.

12. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 11, comprising 78-95% (w/w) THC of the totalphytocannabinoid fraction.

13. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis CBD and the phytocannabinoid containing component comprises 76-96%(w/w) of the cannabis plant extract.

14. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 13, comprising 72-88% (w/w) CBD of the totalphytocannabinoid fraction.

15. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis CBC and the phytocannabinoid containing component comprises 49-60%(w/w) of the cannabis plant extract.

16. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 15, comprising 71-87% (w/w) CBC of the totalphytocannabinoid fraction.

17. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 16, which further comprises the secondaryphytocannabinoids CBD and CBL.

18. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 17, wherein the CBD comprises 6.5-8% (w/w) of thetotal phytocannabinoid fraction and the CBL comprises 5.8-7.1 (w/w) ofthe total phytocannabinoid fraction.

19. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis THCV and the phytocannabinoid containing component comprises 74-90%(w/w) of the cannabis plant extract.

20. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 19, comprising 71-87% (w/w) THCV of the totalphytocannabinoid fraction.

21. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 20, which further comprises the secondaryphytocannabinoid THC.

22. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 21, wherein the THC comprises 14.8-18% (w/w) of thetotal phytocannabinoid fraction.

23. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis CBDV and the phytocannabinoid containing component comprises 64-78%(w/w) of the cannabis plant extract.

24. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 23, comprising 52-64% (w/w) CBDV of the totalphytocannabinoid fraction.

25. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 24, which further comprises the secondaryphytocannabinoids CBD and CBCV.

26. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 25, wherein the CBD comprises 22.4-27.4% (w/w) ofthe total phytocannabinoid fraction and the CBCV comprises 5.5-6.7 (w/w)of the total phytocannabinoid fraction.

27. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis CBGV and the phytocannabinoid containing component comprises 54-66%(w/w) of the cannabis plant extract.

28. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 27, comprising 68-84% (w/w) CBGV of the totalphytocannabinoid fraction.

29. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 28, which further comprises the secondaryphytocannabinoid CBG.

30. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 29, wherein the CBG comprises 19-23% (w/w) of thetotal phytocannabinoid fraction.

31. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis THCA and the phytocannabinoid containing component comprises 54-66%(w/w) of the cannabis plant extract.

32. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 31, comprising 71-86% (w/w) THCA of the totalphytocannabinoid fraction.

33. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 32, which further comprises the secondaryphytocannabinoid THC.

34. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 33, wherein the THC comprises 13.4-16.4% (w/w) ofthe total phytocannabinoid fraction.

35. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis CBDA and the phytocannabinoid containing component comprises 71-86%(w/w) of the cannabis plant extract.

36. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 35, comprising 78-86% (w/w) CBDA of the totalphytocannabinoid fraction.

37. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 36, which further comprises the secondaryphytocannabinoid CBD.

38. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 37, wherein the CBD comprises 6.1-7.5% (w/w) of thetotal phytocannabinoid fraction.

39. A cannabis plant extract or the use of a cannabis plant extract asclaimed in any of paragraphs 1-8, wherein the principle phytocannabinoidis THCVA and the phytocannabinoid containing component comprises 62-75%(w/w) of the cannabis plant extract.

40. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 39, comprising 53-65% (w/w) THCVA of the totalphytocannabinoid fraction.

41. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 40, which further comprises the secondaryphytocannabinoid THCV.

42. A cannabis plant extract or the use of a cannabis plant extract asclaimed in paragraph 41, wherein the THCV comprises 17.3-21.2% (w/w) ofthe total phytocannabinoid fraction.

43. A method of treating a patient comprising administering atherapeutically effective amount of a cannabis plant extract comprisinga phytocannabinoid containing component and a non-phytocannabinoidcontaining component, wherein the phytocannabinoid containing componentcomprises at least 50% (w/w) of the cannabis plant extract and thenon-phytocannabinoid containing component comprises a monoterpenefraction and a sesquiterpene fraction, in which a principle monoterpenesub-fraction is selected from myrcenes or pinenes and a principlesesquiterpene sub-fraction is selected from caryophyllenes or humulenesto the patient.

44. The use of one or more phytocannabinoids, either in an isolated formor in the form of a botanical drug substance (BDS), as a prophylactic orin the treatment of cancer

45. One or more phytocannabinoids, selected from the group consistingof: THCV, CBDV, THCVA, THCA, CBDA, CBD, CBG, and CBC, for use in thetreatment of prostate cancer, wherein, where present, the THCVA ispresent as an isolated phytocannabinoid, the THCA, CBDA, CBD, CBG or CBCare present in the form of a BDS, and the THCV or CBDV are present ineither an isolated form or in the form of a BDS.

46. The use of one or more phytocannabinoids, selected from the groupconsisting of: THCV, CBDV, THCVA, THCA, CBDA, CBD, CBG, and CBC, for usein the manufacture of a medicament to treat of prostate cancer, wherein,where present, the THCVA is present as an isolated phytocannabinoid, theTHCA, CBDA, CBD, CBG or CBC are present in the form of a BDS, and theTHCV or CBDV are present in either an isolated form or in the form of aBDS.

47. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 45 or 46, wherein the one ormore phytocannabinoids are propyl variant phytocannabinoids.

48. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 45 or 46, wherein the one ormore phytocannabinoids are in an acid form.

49. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 45 or 46, wherein the one ormore phytocannabinoids are in a neutral or decarboxylated form.

50. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 45 or 46, wherein thephytocannabinoid is CBG and is in the form of a BDS.

51. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in any of the paragraphs 45 to 50, whereinthe prostate cancer is hormone-sensitive prostate cancer.

52. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 51, wherein thephytocannabinoid is THCVA in an isolated form.

53. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in any of the paragraphs 45 to 50, whereinthe prostate cancer is hormone-insensitive prostate cancer.

54. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 53, wherein thephytocannabinoid is CBD and is in the form of a BDS.

55. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 53, wherein thephytocannabinoid is CBDV and is in the form of a BDS.

56. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 45 or 46, wherein the one ormore phytocannabinoids are used in combination or as an adjunct therapywith a chemotherapeutic agent and/or an anti-androgen.

57. The use as claimed in paragraph 56, wherein the chemotherapeuticagent is a mitotic inhibitor.

58. The use as claimed in paragraph 57, wherein the mitotic inhibitor isfrom the taxane drug class.

59. The use as claimed in paragraph 58, wherein the mitotic inhibitorfrom the taxane drug class is taken from the group: docetaxel;larotaxel; ortataxel; paclitaxel; and tesetaxel.

60. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 56, wherein thephytocannabinoid is CBG.

61. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 56, wherein thephytocannabinoid is CBD.

62. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 60 or 61, wherein thephytocannabinoid is in the form of a BDS.

63. The one or more phytocannabinoids or the use of one or morephytocannabinoids as claimed in paragraph 62, for the purpose of slowingdown the growth or reducing the volume of a prostate cancer tumour.

64. A method of treating a patient with prostate cancer comprisingadministering an effective amount of one or more phytocannabinoids,selected from the group consisting of: THCV, CBDV, THCVA, THCA, CBDA,CBD, CBG, and CBC, wherein, where present, the THCVA is present as anisolated phytocannabinoid, the THCA, CBDA, CBD, CBG or CBC are presentin the form of a BDS, and the THCV or CBDV are present in either anisolated form or in the form of a BDS to the patient.

65. One or more propyl phytocannabinoids or acid phytocannabinoids foruse in the down regulation of ERK signalling and effect one or more of:anti-proliferation, anti-metastasis or anti-angiogenesis in a humanpatient.

66. The use of one or more propyl phytocannabinoids or acidphytocannabinoids in the manufacture of a medicament to down regulateERK signalling and effect one or more of: anti-proliferation,anti-metastasis or anti-angiogenesis in a human patient.

67. The one or more propyl or acid phytocannabinoids or the use of oneor more propyl or acid phytocannabinoids as claimed in paragraph 65 or66, wherein the phytocannabinoids is selected from the group consistingof: THCV, CBGV, CBDV, CBGA and CBDA.

68. The one or more propyl or acid phytocannabinoids or the use of oneor more propyl or acid phytocannabinoids as claimed in any of paragraphs65-67, wherein the phytocannabinoid is in an isolated form.

69. The one or more propyl or acid phytocannabinoids or the use of oneor more propyl or acid phytocannabinoids as claimed in any of paragraphs65-68, for use in the treatment of lung cancer, prostate cancer, orbreast cancer.

70. The one or more propyl or acid phytocannabinoids or the use of oneor more propyl or acid phytocannabinoids as claimed in paragraph 69, foruse in the treatment of bone or lymph metastasis.

71. A method of treating a patient with cancer comprising administeringone or more propyl phytocannabinoids or acid phytocannabinoids to downregulate ERK signalling and effect one or more of: anti-proliferation,anti-metastasis or anti-angiogenesis to the patient.

72. One or more phytocannabinoid acids, excluding CBDA or CBDVA, for usein medicine.

73. One or more phytocannabinoid acids for use in the treatment ofcancer.

74. The use of one or more phytocannabinoid acids in the manufacture ofa medicament for use in the treatment of cancer.

75. One or more phytocannabinoid acids or the use of one or morephytocannabinoid acids as claimed in any of paragraphs 72 to 74, whereinthe one or more phytocannabinoid acid are in the form of a BDS.

76. One or more phytocannabinoid acids or the use of one or morephytocannabinoid acids as claimed in paragraphs 72-75, wherein thecancer to be treated is a cancer of the prostate, breast, colon, lung,glioma or skin.

77. One or more phytocannabinoid acids or the use of one or morephytocannabinoid acids as claimed in paragraphs 72-76, wherein thephytocannabinoid acid is taken from the group consisting of: THCA, CBGAand CBDA.

78. One or more phytocannabinoid acids or the use of one or morephytocannabinoid acids as claimed in paragraphs 72-77, comprising incombination the phytocannabinoid THCA with CBDA and/or CBGA.

79. A method of treating cancer comprising administering a therapeuticamount of one or more phytocannabinoid acids to a patient.

80. An isolated CBD, CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS for usein the treatment of a pre-cancerous symptom of colon cancer.

81. The use of isolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDSin the manufacture of a medicament for use to treat a pre-canceroussymptom of colon cancer.

82. Isolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS or use ofisolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS as claimed inparagraph 80 or 81, wherein the isolated CBD CBG, CBDV, CBD BDS, CBG BDSand/or CBDV BDS are used in the treatment of aberrant crypts in thecolon.

83. Isolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS or use ofisolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS as claimed inparagraph 80 or 81, wherein the isolated CBD CBG, CBDV, CBD BDS, CBG BDSand/or CBDV BDS are used in the treatment of colon polyps.

84. A method of treating a patient with a pre-cancerous symptom of coloncancer, comprising administering a therapeutically effective amount ofan isolated CBD CBG, CBDV, CBD BDS, CBG BDS and/or CBDV BDS to thepatient.

85. A combination of phytocannabinoids together with a chemotherapeuticagent which is not a cannabinoid, for use in the treatment of a glioma.

86. The use of a combination of phytocannabinoids together with achemotherapeutic agent which is not a cannabinoid, in the manufacture ofa medicament to treat a glioma.

87. The combination of phytocannabinoids or the use of the combinationof phytocannabinoids as claimed in paragraph 85 or paragraph 86, whereinthe combination of phytocannabinoids and the chemotherapeutic agentwhich is not a cannabinoid are packaged for administration separately,simultaneously or sequentially.

88. The combination of phytocannabinoids or the use of the combinationof phytocannabinoids as paragraphed in paragraphs 85-87, wherein thephytocannabinoids are THC and CBD.

89. The combination of phytocannabinoids or the use of the combinationof phytocannabinoids as claimed in paragraphs 85-88, wherein the doselevel of the phytocannabinoids is sub-effective for the treatment of theglioma if used alone.

90. The combination of phytocannabinoids or the use of the combinationof phytocannabinoids as claimed in paragraphs 85-89, wherein thechemotherapeutic agent is temazolamide.

91. The combination of phytocannabinoids or the use of the combinationof phytocannabinoids as claimed in paragraphs 85-90, wherein the doselevel of the temazolamide is sub-effective for the treatment of theglioma if used alone.

92. A method of treating a patient with a glioma, comprisingadministering a therapeutically effective amount of a combination ofphytocannabinoids together with a chemotherapeutic agent which is not acannabinoid, to the patient.

The invention claimed is:
 1. A method of treating a human suffering froma glioma consisting essentially of administering therapeuticallyeffective amounts of isolated or highly purified tetrahydrocannabinoland cannabidiol, together with temozolomide to said human to treat saidglioma in said human.